Table 4.

Singleton categorization according to phenotypical impact.

Category	Yield	Gene Transfer	Isolates (Mapped AAV8 VP1 location)
I	+	n/a	rh.64R1(697), hu.44R3 (137, 448, 611), hu.48R2 (324, 279), hu.29R (399)
II	+	+	hu.48R3 (554), rh.48.1 (305), rh.48.2 (217) rh.48R2 (217, 305), rh.37R2 (212)
III	=	+	AAV6.2 (129), cy.5R1(13), cy.5R5 (159+166)
IV	+	=	rh.64R2(686), rh.37R1(643)
V	=	=	rh.2R (651), cy.5R3(51), rh.8R(533)
VI	=	−	cy.5R2 (410)
VII	+	−	AAV6.1 (533), AAV6.1.2 (533)
VIII	no rescue	n/a	hu.46R1–R4, rh.67R, rh.58R2

The effect of a particular singleton reversion on the capsid is categorized according to the effect on vector yield in production and in vivo gene transfer (upon i.v. administration) (+, increase; = no effect; −, decrease). Residues that upon the change demonstrated the effect are mapped onto the homologous AAV8 sequence (VP1 numbering). Residue locations in italics refer to preceding modifications made in the sequential mutagenesis strategy but for which the phenotypical impact could not be attributed directly. n/a: not available